The Pharma Innovation Journal 2021; 10(4): 671-674
ISSN (E): 2277- 7695
ISSN (P): 2349-8242
NAAS Rating: 5.23
TPI 2021; 10(4): 671-674
© 2021 TPI
www.thepharmajournal.com
Received: 16-02-2021
Accepted: 18-03-2021
Dr. Manoj Bhanukar
Ph.D. Scholar Department of
Horticulture, CCS HAU, Hisar,
Haryana, India
Dr. GS Rana
Principal Scientist, Department
of Horticulture, CCS HAU,
Hisar, Haryana, India
Preeti
Post Graduate from Department
of Horticulture, CCS HAU,
Hisar, Haryana, India
Corresponding Author:
Preeti
Post Graduate from Department
of Horticulture, CCS HAU,
Hisar, Haryana, India
Effect of foliar application of micronutrients and soil
application of zinc sulphate on total chlorophyll and
carotenoid content in sweet orange (Citrus sinensis) cv.
blood red
Dr. Manoj Bhanukar, Dr. GS Rana and Preeti
DOI: https://doi.org/10.22271/tpi.2021.v10.i4j.6034
Abstract
The present investigation was carried out for two consecutive years of 2016 and 2017 at experimental
orchard of Department of Horticulture, CCS HAU, Hisar to assess the performance of sweet orange cv.
Blood Red to foliar application of micronutrients and soil application of zinc sulphate. In experiment I,
the foliar application of micronutrients was done two times i.e. Ist spray was done on first week of april
and 2nd spray on first week of july. The experiment comprises of thirteen treatments, viz., Zinc sulphate
(ZnSO4) @ 0.25%, 0.50% &1.00%; Iron sulphate (FeSO4) @ 1.5%, 2.0% & 2.5%; Manganese sulphate
(MnSO4) @ 0.25%, 0.50% & 1.00%; Boric acid (H3BO3) @ 0.1%, 0.2% & 0.3% and Control (No
micronutrients). In experiment II, the soil application of zinc sulphate (ZnSO 4) was done two times i.e.
half dose in second fortnight of february and half dose in second fortnight of May. The experiment
comprises of six treatments, viz., Zinc sulphate (ZnSO4) @ 50 g/plant, 100 g/plant, 150 g/plant, 200
g/plant, 250 g/plant and Control (No Zn application). Results indicated that foliar application of
micronutrients and soil application of zinc sulphate affected significantly the chlorophyll and carotenoid
content.
Keywords: Citrus sinensis, foliar application, soil application, micronutrients, sweet orange
Introduction
The genus Citrus is economically very important and is known for its juice and pulp
throughout the world. Citrus is one of the choicest fruit with high consumer preference both as
fresh fruit as well as for its refreshing processed juice. It is extensively grown in tropical and
sub-tropical regions. Citrus is a commercially important fruit crop of India and grown across
its length and breadth with a production of 13200 thousand MT from an area of 1034 thousand
hectares during year of 2018-19 (Anonymous, 2018-19) [1]. Sweet orange (Citrus sinensis (L.)
Osbeck) has been reported to be originated in Southern China and it was introduced to India
during thirteenth century (Swingle, 1943; Webber, 1948) [9, 11]. It is the second largest citrus
fruit, cultivated in tropical and subtropical regions of the country. Total sweet orange
production in india is 3401 thousand MT with an area of 190 thousand hectares during year of
2018-19 (Anonymous, 2018-19) [1]. The productivity of sweet orange mainly depends on
adequate supply of plant nutrients seems to be a very important factor in regulating cropping
and influencing the quality of sweet orange. The micronutrients are required in small amounts,
but play a great role in plant metabolism (Kazi et al., 2012) [4]. Despite some shortcomings,
foliar application is regarded as the best method under certain conditions (Marschner and
Marschner, 2012) [6]. Foliar or soil application of Zn increases the biosynthesis of chlorophyll
and carotenoid synthesis that are important for proper performance of photosynthetic process
(Mousavi, 2011) [7]. The beneficial effects of amelioration of zinc, iron, manganese and boron
deficiencies by foliar application have been documented by previous studies (Hippler et al.,
2015) [3]. The present investigation has been undertaken to study the effect of foliar application
of micronutrients and soil application of zinc sulphate on chlorophyll and carotenoid content
of fruit sweet orange.
Material and Methods
The present investigation was carried out on fifteen years old sweet orange cv. Blood Red trees
planted at a spacing of 6m X 6m in experimental orchard of Department of Horticulture, CCS
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HAU, Hisar situated at 215.2 m above mean sea level with
coordinates of 29º10ʹ N latitude and 75º46ʹ E longitudes,
during the year 2016-17 and 2017-18. In Experiment I, the
foliar application of micronutrients was done two times i.e. Ist
spray was done on first week of april and 2 nd spray on first
week of july. The experiment comprises of thirteen
treatments, viz., Zinc sulphate (ZnSO4) @ 0.25%, 0.50%
&1.00%; Iron sulphate (FeSO4) @ 1.5%, 2.0% & 2.5%;
Manganese sulphate (MnSO4) @ 0.25%, 0.50% & 1.00%;
Boric acid (H3BO3) @ 0.1%, 0.2% & 0.3% and Control (No
micronutrients). In experiment II, the soil application of zinc
sulphate (ZnSO4) was done two times i.e. half dose in second
fortnight of february and half dose in second fortnight of
May. The experiment comprises of six treatments, viz., Zinc
sulphate (ZnSO4) @ 50 g/plant, 100 g/plant, 150 g/plant, 200
g/plant, 250 g/plant and Control (No Zn application).
Treatments were allocated in randomized block design (RBD)
with three replications in each treatment. The recommended
standard package of practises and plant protection measures
were adopted to keep the plants in good health. Methods used
to measure the chlorophyll and carotenoid content are as
follows:-
Total chlorophyll and carotenoid Content: Chlorophyll
pigments namely chl. a, chl. b, total chlorophyll and
carotenoids were estimated in the fruits taken from different
sampling stages by the method given by Wellburn (1994) [12]
and expressed in terms of µ moles/g fresh weight of peel
tissue.
Table 1: Effect of foliar application of micronutrients on total chlorophyll content (µ moles/g fr. wt.) of sweet orange cv. Blood Red
Total Chlorophyll content
Treatments
2016
ZnSO4 0.25% 180.86
ZnSO4 0.50% 182.03
ZnSO4 1.00% 183.92
FeSO4 1.5% 185.44
FeSO4 2.00% 187.24
FeSO4 2.5% 189.91
MnSO4 0.25% 176.35
MnSO4 0.50% 177.78
MnSO4 1.00% 179.21
H3BO3 0.1% 172.36
H3BO3 0.2% 173.39
H3BO3 0.3% 174.98
Control 171.12
C.D. at 5% 1.16
The data concerning total chlorophyll content is presented in
Table 1, which indicates that different concentrations of
ZnSO4, FeSO4, MnSO4 and H3BO3 affected total chlorophyll
content significantly during both the years. In year 2016,
maximum (189.91) total chlorophyll content was observed
with FeSO4 2.5% followed by FeSO4 2.00% (187.24) and
minimum (171.12) in control. Similar trend was observed in
the year 2017, showing maximum (189.80) total chlorophyll
content was observed with FeSO4 2.5% followed by FeSO4
2.00% (188.14) and minimum (172.81) in control,
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Extraction of pigments: Two discs (0.28 cm2 area) of the
fruit peel were suspended in a test tube containing 10 ml of
dimethyl sulphoxide (DMSO). The weight of the discs was
recorded before putting them into test tubes. The test tubes
were then placed in an oven at 60oC for four hours to facilitate
the extraction of pigments.
Estimation: After the incubation period for extraction, the
test tubes were cooled to room temperature and the optical
density was read at 454, 645 and 665 nm on a
spectrophotometer (Spectronic-20D). DMSO was used as
blank.
The chlorophyll pigments were calculated by using the
following equations:-
Chlorophyll - a (µg/ml) = 12.19 A665 – 3.45 A645
Chlorophyll - b (µg/ml) = 21.99 A645 – 5.32 A665
Total Chlorophyll (µg/ml) = Chlorophyll a + Chlorophyll b
The quantity of the above pigments were calculated in mg g -1
tissue fresh weight and expressed in µ moles/g tissue fresh
weight by using the following relationship:-
µ moles of Chlorophyll – a = µg chl. a X 1.119
µ moles of Chlorophyll – b = µg chl. b X 1.102
µ moles of Carotenoids = µg carotenoids X 1.809
Results and Discussion
Total Chlorophyll content
2017
182.10
183.71
185.22
186.89
188.14
189.80
177.78
179.12
180.56
174.18
175.01
176.37
172.81
1.12
respectively. The results on total chlorophyll were found
significant with foliar application of micronutrients. The
highest total chlorophyll was observed with FeSO4 2.5%. An
increase in total chlorophyll content with foliar application of
micronutrients might be due to delayed chlorophyll
degradation. The results are in conformation with Shahin et
al. (2010) [8] in apple and Eiada and Al-Hadethi (2013) [2] in
pomegranate. This might also be due to delayed senescent
pigment changes and oppose the ethylene induced loss of
chlorophyll (Trebitsh et al., 1993) [10].
The Pharma Innovation Journal http://www.thepharmajournal.com

Table 2: Effect of foliar application of micronutrients on carotenoid content (µ moles/g fr. wt.) of sweet orange cv. Blood Red
Carotenoid content Carotenoid content
Treatments
2016 2017
ZnSO4 0.25% 246.55 249.67
ZnSO4 0.50% 245.01 247.09
ZnSO4 1.00% 242.89 245.80
FeSO4 1.5% 241.08 243.19
FeSO4 2.00% 238.89 241.98
FeSO4 2.5% 235.78 238.87
MnSO4 0.25% 251.17 253.19
MnSO4 0.50% 249.61 251.65
MnSO4 1.00% 248.02 250.12
H3BO3 0.1% 255.50 257.45
H3BO3 0.2% 254.26 256.56
H3BO3 0.3% 252.60 254.62
Control 257.01 259.01
C.D. at 5% 1.41 1.20

The data presented in Table 2 indicate that different increases the biosynthesis of chlorophyll that is important for
concentrations of ZnSO4, FeSO4, MnSO4 and H3BO3 proper performance of photosynthetic process (Mousavi,
significantly affected carotenoid content of sweet orange cv. 2011) [7].
Blood Red fruits during both the years. In the year 2016, the
highest carotenoid content (257.01) was found in control Table 4: Effect of soil application of zinc sulphate on carotenoid
followed by H3BO3 0.1% (255.50) and lowest (235.78) in content (µ moles/g fr. wt.) of sweet orange cv. Blood Red
FeSO4 2.5%. During the year 2017, highest carotenoid content Carotenoid content Carotenoid content
(259.01) was found in control followed by H 3BO3 0.1% Treatments
2016 2017
(257.45) and lowest (238.87) in FeSO4 2.5%. The results on ZnSO4 50 g/plant 252.14 254.23
content were found significant with foliar application of ZnSO4 100 g/plant 250.54 252.14
micronutrients. The highest carotenoid content was found in ZnSO4 150 g/plant 248.89 250.53
control whereas lowest in FeSO4 2.5%. A decrease in ZnSO4 200 g/plant 245.12 247.91
carotenoid content with foliar application of micronutrients ZnSO4 250 g/plant 243.33 245.03
might be due to inhibition of carotenoid beta-cyptoxanthin Control 254.48 256.11
biosynthesis and its accumulation, which inhibit the C.D. at 5% 1.12 1.78
development of citrus fruit colour and luster. The results are
in conformation with Shahin et al. (2010) [8] in apple and Data given in Table 4 indicated that carotenoid content was
Eiada and Al-Hadethi (2013) [2] in pomegranate. significantly affected by various ZnSO4 concentrations during
both the years. In year 2016, the carotenoid content was found
Table 3: Effect of soil application of zinc sulphate on total maximum (254.48) in control followed by ZnSO4 50g/plant
chlorophyll content (µ moles/g fr. wt.) of sweet orange cv. Blood (252.14) and minimum (243.33) with ZnSO4 250g/plant .In
Red the next year 2017, the carotenoid content was recorded
Total chlorophyll Total chlorophyll maximum (256.11) in control followed by ZnSO4 50g/plant
Treatments content content (254.23) and minimum (245.03) with ZnSO 4 250g/plant .
2016 2017 The results carotenoid content were found significant with
ZnSO4 50 g/plant 176.44 176.92 soil application of zinc sulphate. The highest carotenoid
ZnSO4 100 g/plant 177.98 178.02 content was found in control. An decrease in carotenoid
ZnSO4 150 g/plant 179.76 180.23 content might be because zinc inhibit carotenoid beta-
ZnSO4 200 g/plant 181.48 182.91 cyptoxanthin biosynthesis and its accumulation, which inhibit
ZnSO4 250 g/plant 184.67 185.01 the development of citrus fruit colour and luster. The results
Control 172.78 173.66 are in conformity with Shahin et al. (2010) [8] in apple and
C.D. at 5% 1.54 1.24 Eiada et al. (2013) [3] in pomegranate.

Data presented in Table 3 revealed that total chlorophyll References

content was significantly affected by various ZnSO4
concentrations during both the years. In year 2016, the
maximum total chlorophyll content (184.67) was observed
with ZnSO4 250g/plant followed by ZnSO4 200g/plant
(181.48) and minimum (172.78) in control. In year 2017, the
maximum total chlorophyll content (185.01) was observed
with ZnSO4 250g/plant followed by ZnSO4 200g/plant
(182.91) and minimum (173.66) in control.
The highest total chlorophyll was observed with ZnSO 4
250g/plant. This might be because zinc delayed chlorophyll
degradation. Zinc application enhances the rate of
photochemical reductions and chlorophyll content in
cucumber (Kazemi, 2013) [5]. The soil application of Zn
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4. Kazi SS, Sayed I, Joshi KG. Effect of multi-micronutrient
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The Pharma Innovation Journal http://www.thepharmajournal.com

5. Kazemi M. Application of iron and zinc on growth and

productivity of cucumber. Bull. Environ. Pharmacol. Life
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6. Marschner H, Marschner P. Marschner’s mineral
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nova synthesis of chlorophyllase, a fruit peel. Proceeding
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